牦牛体细胞核移植电融合参数和重构胚胎培养方法的优化

Optimization of fusion parameters and culture methods for yak somatic cell nuclear transfer

为了探索牦牛体细胞核移植胚胎培养方法和体系,试验采用不同的脉冲电压和脉冲时间对牦牛重构胚胎进行融合,采用不同的培养方法对牦牛核移植重构胚胎进行培养,从而优化牦牛体细胞核移植胚胎的培养体系。结果表明：牦牛核移植重构胚胎采用16 V、20μs、2次脉冲融合率最高,为（80.8±3.8）%。采用mSOF培养液、G1/G2培养液、输卵管上皮细胞共培养三种培养体系对牦牛体细胞克隆胚胎的卵裂率影响差异不显著（P〉0.05）,但是G1/G2培养液的囊胚率为（20.0±2.4）%,与输卵管上皮细胞共培养的囊胚率[（19.4±2.3）%]相比差异不显著（P〉0.05）,显著高于mSOF培养液的囊胚率[（13.7±3.8）%]（P〈0.05）。

The aim of the present study was to explore the culture methods and systems of yak somatic cell nuclear transfer embryos. The fusion of yak reconstructed embryos was performed by using different pulse voltages and pulse times. Yak somatic cell nuclear transfer embryos were cultured by using different culture methods. So as to optimize the culture system of yak somatic cell nuclear transfer embryos. The results showed that the fusion rate of yak reconstructed embryos was the highest（80. 8±3. 8） % by using pulse voltages of 16 V,pulse duration of 20 μs and two pulse. When the yak reconstructed embryos were cultured by using mSOF medium,G1/G2 medium and co-culture with oviductal epithelial cells,the effect of three culture systems on the cleavage rate of yak reconstructed embryos was not significant difference（P0. 05）. The blastocyst rate of G1/G2 medium（20. 0±2. 4） % and the blastocyst rate（19. 4±2. 3） % under the oviductal epithelial cells co-culture condition was not also significantly different（P0. 05）,but significantly higher than the blastocyst rate of mSOF medium（13. 7±3. 8） %（P0. 05）.